title: Scaling up in vitro transcription synthesis of RNA standards for competitive quantitative RT-PCR: looking for bigger yields.
abstract: Because the acquisition of in vitro transcription kits, for production of RNA standards, might not be affordable for some small laboratories, the current work describes some alternatives to the commercial Promega protocols. Yields of tested reactions were all higher than the ones reported for the standard Riboprobe kit (1mug RNA/mug template, 1x) and the optimized variant for high yields (5-10x). They were also as good as the ones described for the high RNA production kit RiboMAX (10-20x), exceeding them in approximately 70% of reactions. Our best results, achieved in a 500-mul format, were three to five times higher than the ones reported for the Promega kit.
authors.affiliation: Centro de InmunoEnsayo (CIE), Immunochemistry, Calle 134 y Avenide 25 Playa, Apartado Postal 6653, CP 11600, Ciudad de la Habana, Cuba. idania_gonzalez@yahoo.es
authors.forename: Idania
authors.initials: I
authors.lastname: Gonzalez-Perez
authors.forename: Iria García de la
authors.initials: IG
authors.lastname: Rosa
authors.forename: Anny Armas
authors.initials: AA
authors.lastname: Cayarga
authors.forename: Yenitse Perea
authors.initials: YP
authors.lastname: Hernández
authors.forename: Yaimé Josefina González
authors.initials: YJ
authors.lastname: González
authors.forename: Yoamna Rosario
authors.initials: YR
authors.lastname: Víctores
authors.forename: Santiago
authors.initials: S
authors.lastname: Dueñas-Carrera
mesh_terms: D016174:Hepacivirus / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis* / Q000235:genetics; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D014158:Transcription, Genetic
chemical_list: D012367:RNA, Viral